Apoptosis induced in human osteosarcoma cells is one of the mechanisms for the cytocidal effect of Ad5CMV-p53

Cancer Gene Ther. 1995 Mar;2(1):9-17.

Abstract

Recombinant adenovirus Ad5CMV-p53 induced a strong cytocidal effect in Saos-LM2 cells. This cell line, derived from human osteosarcoma Saos-2 cells, has a homozygous deletion of the p53 gene. By using immunocytochemical and Western blot analyses, we demonstrated that Ad5CMV-p53 effectively infected Saos-LM2 cells at multiplicities of infection of 10 to 50 plaque-forming units/cell and mediated a high-level expression of the exogenous wild-type p53 protein in the cells. Growth of the infected cells was greatly suppressed whereas that of mock- or control virus-infected cells was not. Because wild-type p53 induces apoptosis in certain types of cells, we studied DNA fragmentation in situ in the Ad5CMV-p53-infected Saos-LM2 cells by terminal deoxynucleotidyl transferase assays, which yielded positive nuclear staining. Further analysis of the Ad5CMV-p53-infected Saos-LM2 cells by light and electron microscopy demonstrated that the cells underwent the characteristic morphological changes of apoptosis such as plasma-membrane blebbing, nuclear condensation, and fragmentation. These changes were not observed in mock- or control virus-infected cells. Our results on Saos-LM2 cells indicate that apoptosis induced by Ad5CMV-p53 may be one of the mechanisms underlying the cytocidal effect of Ad5CMV-p53.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenoviruses, Human / genetics*
  • Apoptosis* / genetics
  • Bone Neoplasms / genetics
  • Bone Neoplasms / pathology*
  • Cytomegalovirus / genetics*
  • Gene Deletion
  • Gene Expression Regulation, Viral
  • Genes, p53*
  • Genetic Vectors*
  • Humans
  • Microscopy, Electron
  • Neoplasm Transplantation
  • Osteosarcoma / genetics
  • Osteosarcoma / pathology*
  • Recombinant Fusion Proteins / metabolism*
  • Tumor Cells, Cultured
  • Tumor Suppressor Protein p53 / physiology*

Substances

  • Recombinant Fusion Proteins
  • Tumor Suppressor Protein p53